A method for preparing an infectious, recombinant virus vector comprises the steps of: (a) infecting host cells with a first virus comprising an encapsidation defective adenovirus (edAd), the edAd comprising a first defective virus genome; (b) incubating the infected host cells in a culture medium for a period of time sufficient for producing infectious virus particles; and (c) recovering infectious virus particles secreted into a culture supernatant, wherein the edAd or the host cells comprise a second defective virus genome engineered to express a target gene of interest, wherein the edAd or the host cells comprise nucleic acid sequences sufficient for expressing adenovirus (Ad) helper genes necessary for replication of the defective virus DNA; and wherein the edAd or the host cells comprise nucleic acid sequences sufficient from expressing helper functions necessary for producing infectious, replication defective virus particles corresponding to the second virus.

Described herein are methods for preventing, inhibiting, or treating cancer in a subject. Also provided herein are methods of altering expression of one or more gene products in a cell, such as a cancer cell. Such methods may comprise utilizing a modified nuclease system, such as Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR associated (Cas) 9 (CRISPR-Cas9) comprising a bidirectional HI promoter and gRNAs directed to oncogenes (rAAV-Onco-CRISPR) or tumor suppressor genes (rAAV-TSG) packaged in a compact adeno-associated virus (AAV) particle. Such methods may comprise co-administering or concurrently providing a recombinant adeno-associated virus-packaging adenovirus (Ad-rAAVpack) with the nuclease system.

The presently disclosed subject matter provides compositions and methods comprising improvements of a CRISPR system (e.g. CRISPR associated (Cas) 9 (CRISPR-Cas9, non-Cas9 CRISPR systems). Such compositions may comprise modifications to the H1 promoter region, addition of 5UTR modifications, different orthologous sequences of the H1 promoter, novel compact bidirectional promoter sequences with both pol II and pol III activity, addition of Kozak consensus sequences, termination sequences, addition of conditional pol II/pol III bidirectional promoter expression, addition of a donor template sequence for correcting mutations, or combinations thereof. Other aspects of the invention relate to modifications to Cas9 through post-transcriptional cell-cycle regulation fusions, engineered partial target sites such that the nuclease can bind without DNA cleavage, auto-regulation sites, and N-terminal modifications to modulate half-life.

The present disclosure provides, among other things, Ad35 helper genomes and vectors useful in gene therapy, e.g., for production of helper-dependent Ad35 donor vectors. Helper genomes of the present disclosure include a conditionally defective packaging sequence.

The present disclosure provides improved adenoviral helper plasmids for the production of recombinant adeno-associated viruses.

Described is an adenovirus derived helper virus which may comprise the adenoviral DNA sequences for E2a, S4 (orf6), the VA1 RNA gene, and the parvovirus VP capsid gene unit. Also described is a method of efficiently preparing a recombinant parvovirus (particle) which is based on the use of various adenoviral derived helper viruses/vectors.

Described is an adenovirus derived helper virus which may comprise the adenoviral DNA sequences for E2a, S4 (orf6), the VA1 RNA gene, and the parvovirus VP capsid gene unit. Also described is a method of efficiently preparing a recombinant parvovirus (particle) which is based on the use of various adenoviral derived helper viruses/vectors.

Provided is an oncolytic virus expressing interferon. Specifically provided is an oncolytic adenovirus comprising a fusion protein expressing interferon, said oncolytic virus being capable of effectively suppressing tumors in vitro and in vivo.

The present invention is a chimeric adenovirus vector in which the knob domain of the end of the fiber protein of human adenovirus type 5 is replaced with the knob gene of chimpanzee adenovirus serotype 6 and/or in addition the hexon protein of human adenovirus type 5 is replaced with hypervariable regions 1-7 of human adenovirus serotype 28. The present invention not only provides the optimal adenovirus vector in the development of treatments or vaccines for various diseases, but also when the chimeric adenovirus vector produced in the present invention is infected with a host cell for production, it can contribute to improved productivity by exhibiting superior cell infection ability compared to the recombinant HAdV-5 vector-based vaccine.

Provided herein are engineered transcription factors for selective upregulation of SCN1a and uses thereof for treating diseases and disorders, such as, Dravet syndrome. Also provided are microRNA binding sites and uses thereof for selective expression in parvalbumin neurons.